We obtained Panamanian golden frogs (Atelopus varius) from the Maryland zoo as part of the captive breeding program 'Project Golden Frog'. For each frog, we measured mass to the nearest 0.01 g, and snout-vent length (SVL) to the nearest 0.1 mm using calipers and scales. For experiment one, we placed frogs in either high RH (relative humidity), medium RH, or low RH conditions. We collected skin swab samples to test for Bd presence and infection intensity using standardized swabbing techniques. To collect frog skin secretions, we induced half of the frogs from each treatment group to secrete their store of secretions from cutaneous granular glands by administering a stimulatory injection of 40 nmol/gram of norepinephrine (NE) below the skin surface (Rollins-Smith et al. 2005, Woodhams et al. 2006). The remaining frogs from each RH group were given shots of sterile saline as a control. Following a month at each humidity treatment, we injected frogs once again with either norepinephrine or saline, and then returned them to their humidity chambers. Immediately following this second reduction, we exposed frogs to the Bd isolate Rio Maria. We measured mass and SVL weekly to calculate frog body condition (mass/SVL). We swabbed frogs weekly and used a quantitative polymerase chain reaction (qPCR) assay to assess the prevalence and intensity of Bd infection in our diagnostic samples. We analyzed the quantity of skin secretions collected from each group of frogs using a MicroBCA assay using bradykinin to generate a standard curve. We assessed the inhibitory effects of these skin secretions against Bd in vitro using an MTT assay (Lindauer et al. 2019).

For the second experiment, we placed frogs in one of the following conditions: high RH and high temperature, high RH and low temperature, low RH and high temperature, or low RH and low temperature. We exposed half of the frogs from each chamber to the Bd isolate Rio Maria. We measured mass and SVL weekly to calculate frog body condition (mass/SVL). We swabbed frogs weekly and used a quantitative polymerase chain reaction (qPCR) assay to assess the prevalence and intensity of Bd infection in our diagnostic samples.

Data are in .csv files with sheets:

1. Peptide_recovery_data

Frog_ID: individual frog identification
Group: the humidity treatment assigned to each frog
Protein: the amount of skin secretions collected at timepoint 1 in micrograms
Protein 2: the amount of skin secretions collected at timepoint 2 in micrograms

2. Humidity_Bd_load

Frog_ID: individual frog identification
Bd_load: the Bd load (genomic equivalents) for each frog at that timepoint. We used a quantitative measure of Bd genomic equivalents using a real-time PCR assay with universal Bd standards (0.1-100 zoospore equivalents). 
Timepoint: the week that the Bd load was recorded
Date: the calendar date that corresponds with each week
Group: the humidity treatment assigned to each frog
Exposure_group: whether the frogs were exposed to Bd or a saline control
PLUS 1: Adds ‘+1’ to the Bd load column (allows us to take the log of the Bd load).
Log_GE: the log of the Bd load + 1 for each frog

3. Environmental_con_Bd_load

Frog_ID: individual frog identification
Humidity_group: the humidity and temperature treatment that each frog was assigned
Exposure: whether the frog was exposed to Bd or was a control frog.
Timepoint: The week that the Bd load was recorded
Date: The calendar date that corresponds with each week
Bd_load: the Bd load (genomic equivalents) for each frog at that timepoint. We used a quantitative measure of Bd genomic equivalents using a real-time PCR assay with universal Bd standards (0.1-100 zoospore equivalents). 
Plus_1: the Bd load column plus 1 (allows us to take the log of Bd load)
Log_GE: the log of the Bd load +1 for each frog

4. Humidity_survival

Frog_ID: individual frog identification
DOD: the calendar date that each frog died (DOD: date of death)
Group: the humidity treatment group that each frog was placed in
Days survived: the number of days each frog survived

5. Environmental_con_survival

Frog_ID: individual frog identification
DOD: the calendar date that each frog died (DOD: date of death)
Group: the humidity and temperature group that each frog was placed in
Days survived: the number of days each frog survived
Week: the number of weeks each frog survived

6. Inhibitory_effectiveness

Frog_ID: individual frog identification
Group: the humidity treatment group of each frog
Timepoint_1: the inhibitory effect of peptides against Bd from frogs taken at timepoint 1. Inhibitory effectiveness of the peptides is based on their ability to limit Bd growth. The amount of Bd present after addition of the peptides is measured by optical density (OD 570 nm). 
Timepoint_2: the inhibitory effect of peptides against Bd from frogs taken at timepoint 2. Inhibitory effectiveness of peptides is based on their ability to limit Bd growth. The amount of Bd present after addition of the peptides is measured by optical density (OD 570 nm). 

7. Humidity_Body_condition

Frog_ID: individual frog identification
Treatment: The humidity group (high, medium, low) and exposure status (Bd exposed or control) of each frog
Time: The week that the body condition was recorded
Date: The calendar date corresponding to each week
Mass: The mass of each frog at each timepoint measured in grams (g)
SVL: The SVL (Snout-Vent-Length) of each frog at each timepoint measured in millimeters (mm)
Body condition: The mass divided by the SVL for each frog at each timepoint

8. Environmental_con_body_con

Frog_ID: individual frog identification
Treatment: The humidity/temperature group and exposure status (Bd exposed or control) of each frog.
Time: The week that body condition was recorded
Date: The calendar date corresponding to each week
Mass: The mass of each frog at each timepoint measured in grams (g)
SVL: The SVL (Snout-Vent-Length) of each frog at each timepoint measured in millimeters (mm)
Body condition: The mass divided by the SVL for each frog at each timepoint

9. Panama_env_data

Date Time: The date and time that the temperature and humidity was recorded
Ttemp: The temperature (degrees Celsius) taken at the first field station
TRH: The relative humidity taken at the first field station
Btemp: The temperature (degrees Celsius) taken at the second field station
BRH: The RH taken at the second field station

10. Lab_temp_hum

Date: The date and time that temperature and humidity data were collected
Chamber 1 Temp, °F: The temperature from chamber 1 (High RH High Temp)
Chamber 1 RH, %: The Relative humidity from chamber 1 (High RH High Temp)
Chamber 2 Temp, °F: The temperature from chamber 2 (High RH Low Temp)
Chamber 2 RH, %: The relative humidity from chamber 2 (High RH Low Temp)
Chamber 3 Temp, °F: The temperature from chamber 3 (Low RH High Temp)
Chamber 3 RH, %: The relative humidity from chamber 3 (Low RH High Temp)
Room Temp, °F: The temperature from the room (Low RH Low Temp)
Room RH, %: The relative humidity from the room (Low RH Low Temp)